Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound   Target Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound

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Query: EC:3.1.30.2 (endonuclease)
18,621 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The structural gene for the major outer membrane protein (MOMP) from Chlamydia trachomatis was cloned and sequenced. A lambda gt11 recombinant (lambda gt11/L2/33) that contains a portion of the MOMP coding sequence was used to probe a lambda 1059 library constructed from DNA obtained from C. trachomatis serovar L2. Selected lambda 1059 recombinants were mapped with endonuclease restriction enzymes. The MOMP gene was mapped to the 5' end of a BamHI fragment of approximately 9 kilobases. Contiguous endonuclease restriction fragments identified within this region permitted the selection of specific fragments for subcloning and DNA sequencing. The MOMP gene consisted of a 1,182-base-pair open reading frame that encoded 394 amino acids and ended with three stop codons. The known amino-terminal amino acid was preceded by 22 amino acids whose sequence was compatible with a leader or signal sequence. The primary structure of MOMP determined from the translated DNA sequence demonstrated nine cysteine residues and a remarkably homogeneous distribution of charged and hydrophobic residues.
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PMID:Sequence analysis of the major outer membrane protein gene from Chlamydia trachomatis serovar L2. 294 65

Plasmid DNA from an avian strain of Chlamydia psittaci was purified and estimated to be 7.9 kb in size using restriction endonuclease analysis. A 5.9 kb fragment of this plasmid was cloned, mapped and used to screen a range of chlamydial strains. Hybridizing DNA was absent from ovine abortion and arthritis isolates and also from the Cal 10 strain but related sequences were detected in C. psittaci strains of feline pneumonitis, guinea-pig inclusion conjunctivitis, ovine conjunctivitis and C. trachomatis serovar L2. The plasmid DNA from the feline strain was shown to have a distinct restriction endonuclease profile. Similar plasmid sequences were detected in all avian isolates tested: thus the clone may have a useful diagnostic role for the detection of the pathogen in its natural host and in zoonotic episodes.
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PMID:Distribution of plasmid sequences in avian and mammalian strains of Chlamydia psittaci. 318 16

The gene encoding an 18,000-dalton eucaryotic cell-binding protein of Chlamydia trachomatis serovar L2 was cloned into Escherichia coli, and the nucleotide sequence of a 1,658-base-pair PstI restriction endonuclease fragment encoding this protein was determined. The recombinant chlamydial gene consists of a 486-base-pair open reading frame encoding a polypeptide of molecular weight 18,314. The resultant polypeptide, comprising 162 amino acids, possesses a highly charged carboxy-terminal end. The expression of this recombinant protein is under the control of a vector promoter. The recombinant 18,000-dalton protein possessed the same eucaryotic cell-binding characteristics as did the native chlamydial 18,000-dalton protein when electrophoresed and transferred to nitrocellulose. Polyclonal antibodies to the recombinant protein exhibited neutralizing activity.
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PMID:Cloning, expression, and primary structure of a Chlamydia trachomatis binding protein. 331 67

Plasmids from Chlamydia trachomatis LGV-434 (serotype L2) and Chlamydia psittaci meningopneumonitis strain Cal-10 were cloned into the BamHI and EcoRI sites of pBR322, respectively. The recombinant plasmids pCTL2 and pCPMn, each containing an entire respective chlamydial plasmid, were transformed into Escherichia coli. The sizes of the plasmids of C. trachomatis and C. psittaci were 7.3 and 6.2 kilobases, respectively. The two plasmids were found to be distinct by restriction endonuclease analysis, DNA-DNA hybridization, and electron microscopic heteroduplex analysis. However, partial homology was observed between restriction fragments of pCTL2 and pCPMn by Southern blot analysis. Polypeptide products encoded by these plasmids were synthesized in vitro by an E. coli-directed transcription-translation system and in vivo in E. coli maxicells and minicells. None of these polypeptides was immunoreactive with anti-chlamydial sera by immunoblotting or immunoprecipitation. Based on the comparative analysis data, the C. trachomatis and C. psittaci plasmids were found to share little genetic relatedness.
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PMID:Molecular characterization of Chlamydia trachomatis and Chlamydia psittaci plasmids. 394 8

Preparations of DNA from 12 Chlamydia psittaci isolates and one Chlamydia trachomatis strain were compared by restriction endonuclease analysis. Polyacrylamide gel electrophoresis, followed by silver staining, resulted in optimal resolution of fragments generated by digestion. By this technique, four distinct electropherotypes were demonstrated when ovine abortion, ovine arthritis, and avian and Cal10 strains of C. psittaci were examined. Minor profile differences allowed the discrimination of avian isolates derived from psittacine and columbiforme species, and the Cal10 DNA electropherotype was shown to have features in common with these profiles. However, there were no detectable differences in the DNA patterns of eight ovine abortion isolates.
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PMID:Comparison of Chlamydia psittaci isolates by DNA restriction endonuclease analysis. 608 26

The DNA from six serovars of Chlamydia trachomatis, lymphogranuloma venereum (LGV) I, LGV II, LGV III, B, C, and D, and from Chlamydia psittaci was extracted, treated with restriction endonuclease enzymes, and run on agarose gels. By using this technique, the DNA of C. trachomatis could be clearly differentiated from C. psittaci DNA. A comparison of the DNA from the different serovars of C. trachomatis revealed similar patterns with and without detectable differences. LGV I, LGV II, LGV III, B, and C revealed no differences when treated with BamHI, HaeIII, XbaI, and XhoI. LGV III DNA, when cleaved with EcoRI and HhaI, had a major band migrating faster than the other two LGV serovars. Serovar D had a different pattern from all other strains tested when cleaved with BamHI, EcoRI, HhaI, HincI, and XhoI. When treated with SacI and HgaI, LGV II displayed a unique band not seen in the other LGV serovars. Differences in strains could be attributed to both chromosomal and plasmid DNA.
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PMID:Characterization of Chlamydia DNA by restriction endonuclease cleavage. 630 76

The application of a diagnostic and genotyping technique based on the polymerase chain reaction (PCR) to the study of trachoma epidemiology in the Gambian village of Jali is reported. PCR based on the major outer membrane protein (MOMP) gene of Chlamydia trachomatis appears to be more sensitive than either isolation or antigen detection by enzyme immunoassay; it had a specificity of 95% and sensitivity of 51% against clinical signs. PCR genotyping identified genotypes A and B of Chlamydia trachomatis circulating in Jali. Sequencing revealed a Pst1 restriction endonuclease site in the amplified MOMP gene of some B strains but not others; Pst1 digestion of the PCR product proved an easy method of distinguishing these strains. The distribution of serotypes and B strain variants shows a significant degree of household clustering (p < 0.001). PCR based genotyping combined with strain typing provides a new and powerful epidemiological tool for the study of transmission events in trachoma.
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PMID:Molecular epidemiology of trachoma in a Gambian village. 784 72

Chlamydia trachomatis serovar was determined by analysis of restriction fragment length polymorphism in the gene encoding the major outer membrane protein (MOMP) from 435 urogenital specimens. Of the specimens, 254 grew < 25 inclusions and 14 were negative in culture. Although previous studies defined serovar by epitopes or sequences representing only the four variable domains in MOMP, restriction endonuclease sites characteristic for each serovar not only within but also outside these variable domains were cataloged in this study. Novel serovars that grew poorly or not at all in vitro were not observed, and all samples proved similar or identical to one of the 15 known serovars. There was no significant difference in proportions of serovars between men and women. In women, F serovars were more frequently observed in infections with few inclusions in culture, whereas B group serovars predominated when many inclusions were observed.
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PMID:Chlamydia trachomatis serovars in 435 urogenital specimens typed by restriction endonuclease analysis of amplified DNA. 810 52

When the present chlamydial classification was established it was recognized that a wide variety of types were contained within the arbitrary designation Chlamydia psittaci. Early workers relied mostly on observations of growth characteristics to differentiate the types of C. psittaci isolated from a wide range of different hosts. The differences between isolates were confirmed serologically using a variety of tests of which the most sensitive was the micro-immunofluorescence (MIF) test which was able to recognize nine immunotypes among the mammalian isolates alone. This approach has recently been improved by the use of monoclonal antibodies in the MIF test which has confirmed most of the mammalian immunotypes and divided the avian strains into four groups. Studies on the nucleic acid of C. psittaci isolates show clear differences in the size distribution of DNA fragments produced by restriction endonuclease digestion of the genomes of the various types. Most importantly, studies of DNA/DNA homologies showed that at least four of the types identified by biological, serological and restriction endonuclease tests were sufficiently different to be considered separate species. Most recently, attention has been focused on DNA sequence comparisons of C. psittaci genes amplified by the polymerase chain reaction (PCR). The usual target has been the major outer membrane protein gene for which much sequence information is now available. The combination of PCR and MIF with monoclonals has provided a set of practical techniques with which all chlamydial isolates can be detected and typed with relative ease. It is likely that these developments will lead to the reclassification of the genus and, hopefully, a rapid increase of our understanding of the diseases caused by C. psittaci.
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PMID:Typing Chlamydia psittaci--a review of methods and recent findings. 829 58

Twelve reference and four Northern Ireland ovine Chlamydia psittaci isolates including ovine abortion, faecal, conjunctivitis and arthritis isolates were compared. Inclusion morphology was shown to provide a useful means of differentiating the abortion and the non-abortion isolates studied. Identical SDS-PAGE polypeptide profiles were produced by the ovine abortion isolates. The polypeptide profiles of the non-abortion isolates were similar to one another and clearly distinct from the abortion isolate profiles. The restriction endonuclease profiles of the abortion isolates were remarkably similar whereas different profiles were produced by most of the non-abortion isolates. Monoclonal antibodies were prepared and characterized. A number of these reacted with all the isolates of chlamydia tested. Three mAbs reacted exclusively with the ovine abortion isolates while four mAbs reacted exclusively with a number of the faecal isolates.
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PMID:Comparison of ovine abortion and non-abortion isolates of Chlamydia psittaci using inclusion morphology, polyacrylamide gel electrophoresis, restriction endonuclease analysis and reactivity with monoclonal antibodies. 836 94


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